Background:Accumulative research is in progress to clarify clinical aspects of GBV-C. The possibility of interaction between HCV and GBV-C as well as its consequence on development of liver diseases is the most important clinical aspect which encourages researchers to develop a rapid and cost effective technique for simultaneous detection of both viruses. Methods: In this study, a SYBR Green real time multiplex RT-PCR technique as a new economical and sensitive method was designed and validated for simultaneous detection of HCV/GBV-C in HCV positive plasma samples. SYBR green real time RT-PCR technique optimization was performed separately for each virus. Multiplex PCR was established next. Standard sera with known concentrations of HCV RNA and dual HCV/GBV-C positive control samples along with negative control samples were used to validate the assay. Results and Conclusions: Fifty six non cirrhotic HCV positive plasma samples [29 of genotype 3a and 27 of genotype 1a] were collected from patients before receiving treatment. 20.6% of genotype 3a and 18.7% of genotype 1a showed HCV/GBV-C co-infection. As a result, 19.6% of 56 samples had HCV/GBV-C co-infection that was compatible with other results from all over the world. SYBR Green real time multiplex RT-PCR technique can be used to detect HCV/GBV-C co-infection in plasma samples. Furthermore, with application of this method more time and cost could be saved in clinical-research settings.

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